No abstract

Yeast tolerance to Cr (III) and Cr (VI) as well as chromium accumulation potential were shown to depend on treatment time, metal concentration, biomass density and the phase of growth. Kinetic studies as exemplified by Pichia guilliermondii ATCC 201911 revealed a biphasic mode of Cr (III) uptake: a rapid sorption phase was followed by a slow process of accumulation, in which the contribution of the cell-bound Cr fraction increased, while the total cellular Cr level remained constant. Cr (VI) uptake was characterized by a time-dependent increase of total Cr and by a constant fractional contribution of the cell-adsorbed chromium, which suggests that the amount of cell-accumulated Cr also tended to increase over time. The resistance to Cr and metal accumulation levels were substantially elevated for a given strain when cultures were treated at high initial biomass densities (1 mg dry weight/ml) of exponentially proliferating cells. Maximum accumulation capabilities ranged between 4.0 and 13 mg Cr (III)/g dry weight and 2-6.7 mg Cr (VI)/g dry weight. The total cell-accumulated Cr contained 29.3% and 52.3% of organically bound chromium for the treatment of P. guilliermondii with Cr (III) and Cr (VI), respectively. Selected yeast strains, under specified physiological conditions, can be applied for bioremediation of environmental Cr contamination, and might be useful too for attempts to obtain chromium-enriched biomass containing biostabilized and nontoxic Cr forms for nutritional applications.

A comparative study has been made on the sensitivity of the yeast Pichia guilliermondii to Cr (III) and Cr (VI) as well as on the Cr uptake potential at growth-inhibitory concentrations of chromium. The strains used in the study were either isolated from natural sources or obtained from a laboratory strain collection. The results show that most of the natural strains were more tolerant to chromium and were able to grow in the presence of 5 mM Cr (III) or 0.5 mM Cr (VI), that is at concentrations which substantially inhibited the growth of laboratory strains. The cellular Cr content after treatment was similar for both strain types and ranged from 1.2-4.0 mg/g d.w. and 0.4-0.9 mg/g d.w., for Cr (III) and Cr (VI) forms, respectively, however, in one case of a natural strain it reached the value of 10 mg Cr (III)/g dry mass. Natural-source strains were grouped into four groups based on the yeasts' differential response to Cr (III) and Cr (VI). Hexavalent Cr-resistant mutants of a P. giuilliermondii laboratory strain, which revealed markedly changed capabilities of chromium accumulation, were obtained by means of UV-induced mutagenesis. Cr (VI) treatment triggered oversynthesis of riboflavin and the addition of exogenous riboflavin increased P. guilliermondii resistance to both Cr (III) and Cr (VI). Electrophoretic protein profiles revealed the induction and/or suppression of several proteins in response to toxic Cr (VI) levels.

This paper reports on the experimental data supporting an essential role of extracellular reduction in chromate detoxification by baker's and non-conventional yeasts. A decrease of chromate content in the yeast culture coincides with an increase of Cr(III) content in extracellular liquid. At these conditions, cell-bound chromium level was insignificant and a dominant part of extra-cellular Cr(III) species was detected in the reaction with chromazurol S only after mineralization of the cell-free samples. This phenomenon of chromium "disappearance" can be explained by the formation of Cr(III) stable complexes with extra-cellular yeast-secreted components which are "inaccessible" in the reaction with chromazurol S without mineralization. It was shown that increasing sucrose concentration in a growth medium resulted in an increase of chromate reduction. A strong inhibition of chromate reduction by 0.25 mM sodium azide, a respiration inhibitor and a protonophore, testifies that extra-cellular chromate detoxification depends on energetic status of the yeast cells. It was shown that Cr(III)-biochelates produced in extra-cellular medium are of a different chemical nature and can be separated into at least two components by ion-exchange chromatography on anionit Dowex 1x10. A total yield of the isolated Cr(III)-biocomplexes is approximately 65 % (from initial level of chromate) with a relative molar ratio 8:5.